CAJ | 학술논문

目的：探讨缝隙连接通讯渐弱剂18α－甘草次酸（ GA）对脂肪细胞向成骨细胞横向分化及细胞间缝隙连接通讯的影响。方法选取3月龄新西兰大白兔腹股沟处脂肪组织，分离提取成熟脂肪细胞，使其去分化得到去分化脂肪细胞，取第三代去分化的脂肪细胞进行成骨诱导，加入GJIC渐弱剂18α－GA设为减弱组，并设立对照组， MTT法观察18α－GA对细胞倍增是否有影响，进行茜素红钙结节染色和I型胶原免疫组化染色对成骨细胞定性检测，Western blot技术和划痕标记染料示踪法检测缝隙连接蛋白43（ Cx43）的表达。结果MTT法检测结果显示18α－GA对细胞生长增殖无显著影响。茜素红染色显示均发现紫红色结节。免疫组化染色显示减弱组I型胶原表达减弱，SLDT 法显示减弱组荧光染料扩散低于对照组，Western blot技术检测结果显示减弱组Cx43蛋白的表达下降。结论18α－GA可以减弱缝隙连接通讯以降低脂肪细胞向成骨细胞分化的能力。
목적：탐토봉극련접통신점약제18α－감초차산（ GA）대지방세포향성골세포횡향분화급세포간봉극련접통신적영향。방법선취3월령신서란대백토복고구처지방조직，분리제취성숙지방세포，사기거분화득도거분화지방세포，취제삼대거분화적지방세포진행성골유도，가입GJIC점약제18α－GA설위감약조，병설립대조조， MTT법관찰18α－GA대세포배증시부유영향，진행천소홍개결절염색화I형효원면역조화염색대성골세포정성검측，Western blot기술화화흔표기염료시종법검측봉극련접단백43（ Cx43）적표체。결과MTT법검측결과현시18α－GA대세포생장증식무현저영향。천소홍염색현시균발현자홍색결절。면역조화염색현시감약조I형효원표체감약，SLDT 법현시감약조형광염료확산저우대조조，Western blot기술검측결과현시감약조Cx43단백적표체하강。결론18α－GA가이감약봉극련접통신이강저지방세포향성골세포분화적능력。
Objective It is to explore the effects of transdifferentiation from adipocytes to osteoblasts and gap junction in-tercellular communication( GJIC) by gap junction Diminuendo agent 18α-GA.Methods Groin adipose tissue was taken from 3 months old New Zealand White Rabbit and collected mature adipocytes.Made the mature adipocytes dedifferentiate and ob-tained the dedifferentiated adipocytes.The 3rd generation of the dedifferentiated adipocytes were induced and differentiated in-to osteoblasts.There were two experimental groups that the diminuendo group with adding 18α-GA and the control group. The toxic action of 18α-GA in osteoblasts growth and proliferation were detected by MTT assay.Osteoblasts were qualitatively detected by alizarin red staining and collagen typeⅠimmunohistochemistry staining.The expressions of Cx43 were measured by western-blot and sorape-loading and dye transfet ( SLDT) method.Results The results of MTT assay showed that 18α-GA had no significant effect on cell proliferation.The results of Alizarin red staining showed that calcific nodules were dyed red in each group.Immunohistochemistry staining results showed that the expressions of collagen typeⅠwere weakened in the diminuendo group.The results of Western blot showed that the expressions of Cx43 were decreased in the diminuendo group. Conclusion 18α-GA diminuendo the capability of transdifferentiation from adipocytes to osteoblasts with enhancement of GJIC.